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Author Notes:

Corresponding authors: Chunhui Xu, PhD, Associate Professor, Department of Pediatrics, Emory University School of Medicine, 2015 Uppergate Drive, Atlanta, Georgia 30322. chunhui.xu@emory.edu or Gang Bao, PhD, Professor, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory Unviersity, 313 Ferst Dr NW, Atlanta, GA. 30332. gang.bao@bme.gatech.edu

Rajneesh Jha and Brian Wile contributed equally to this work.

We thank Aaron Rae at the Emory Children’s Pediatric Research Flow Cytometry Core for assistance with flow cytometry analysis and cell sorting.

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Subjects:

Research Funding:

This study was supported in part by an NIH-NHLBI grant R21HL118454 (C.X. and G.B.) and in part by the NIH-NHLBI Program of Excellence in Nanotechnology award under Contract No. HHSN268201000043C (G.B. and C.X.).

Keywords:

  • Technology
  • Engineering, Biomedical
  • Materials Science, Biomaterials
  • Engineering
  • Materials Science
  • Cardiomyocyte
  • Flow cytometry
  • Gene expression
  • Molecular imaging
  • Stem cell
  • MESSENGER-RNA
  • LIVING CELLS
  • HEART
  • DIFFERENTIATION
  • MYOCYTES

Molecular beacon-based detection and isolation of working-type cardiomyocytes derived from human pluripotent stem cells

Tools:

Journal Title:

Biomaterials

Volume:

Volume 50, Number 1

Publisher:

, Pages 176-185

Type of Work:

Article | Post-print: After Peer Review

Abstract:

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) provide a potential source of cells to repair injured ventricular myocardium. CM differentiation cultures contain non-cardiac cells and CMs of both nodal and working subtypes. Direct application of such cultures in clinical studies could induce arrhythmias; thus, further purification of working-type CMs from heterogeneous cultures is desirable. Here, we designed 10 molecular beacons (MBs) targeting NPPA mRNA, a marker associated with working-type CMs and highly up-regulated during differentiation. We examined these MBs by solution assays and established their specificity using NPPA-overexpressing CHO cells as well as hPSC-CMs. We selected one MB for subsequent CM subtype isolation using fluorescence-activated cell sorting because the signal-to-background ratio was the highest for this MB in solution assays and a linear correlation was observed between MB signals and the CM purity in differentiation cultures. Compared with cells with low MB signals, cells positively selected based on MB signal had higher expression levels of genes associated with working-type CMs and lower expression levels of genes associated with nodal-type CMs. Therefore, the MB-based method is capable of separating working-type CMs from nodal-type CMs with high specificity and throughput, potentially providing working-type CMs for biomedical applications.

Copyright information:

© 2015 Elsevier Ltd.

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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